The Primerdesign genesig Kit for Shigella species (Shigella_spp) genomes is designed for the in vitro quantification of Shigella_spp genomes. The kit is designed to have a broad detection profile. Specifically, the primers represent 100% homology with over 95% of the NCBI database reference sequences available at the time of design.
The dynamics of genetic variation means that new sequence information may become available after the initial design. Primerdesign periodically reviews the detection profiles of our kits and when required releases new versions.
The target sequence is within the virulence plasmid pCP301 (VirA) which is carried by nearly all clinical isolates of shigella. This target has previously been shown to be a good genetic marker for Shigella in other real time PCR based studies (Hiroshi Fukushima et.al 2003.) The primers and probe sequences in this kit have 100% homology with over 95% of reference sequences in the NCBI database based on a comprehensive bioinformatics analysis.
Exceptional value for money
Rapid detection of all clinically relevant subtypes
Positive copy number standard curve for quantification
Highly specific detection profile
High priming efficiency
Broad dynamic detection range (>6 logs)
Sensitive to < 100 copies of target
Accurate controls to confirm findings
This product is specially designed for stool RNA extraction. The kit is suitable for extracting high-purity microbial or host cell RNA from ≤ 0.15g stool samples. The purified RNA can be directly used in RT-PCR and Northern hybridization.
The Kit combines the speed and efficiency of silica-based technology with the convenient handling of magnetic particles for purification of total RNA. Samples are lysed and RNA is purified from lysates in one step through its binding to the silica surface of the particles in the presence of a chaotropic salt. The particles are separated from the lysates using a magnet and DNA is removed by treatment with RNase-free DNase. The magnetic particles are efficiently washed, and RNA is eluted in RNase-free water.
Kit Contents
| Contents | R662601 | R662602 | R662603 |
| Purification Times | 48 Preps | 96 Preps | 5 x 96 Preps |
| MagPure RNA Particles | 1.7 ml | 4.0 ml | 18 ml |
| DNase I | 600 μl | 2 x 600 μl | 10 x 600 μl |
| DNase Buffer | 30 ml | 40 ml | 200 ml |
| Buffer SPL | 30 ml | 60 ml | 270 ml |
| Buffer PHC | 30 ml | 60 ml | 270 ml |
| Buffer MCB* | 9 ml | 15 ml | 75 ml |
| Buffer ALB3* | 10 ml | 20 ml | 100 ml |
| Buffer GW1* | 44 ml | 66 ml | 2 x 220 ml |
| Buffer RW2* | 20 ml | 50 ml | 2 x 100 ml |
| RNase Free Water | 10 ml | 30 ml | 120 ml |
| 2ml Bead Tubes | 48 | 96 | 5 x 96 |
Storage and Stability
MagPure RNA Particles should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, MagPure RNA Particles up to 8 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under these conditions.
This product is specially designed for stool RNA extraction. The kit is suitable for extracting high-purity microbial or host cell RNA from ≤ 0.15g stool samples. The purified RNA can be directly used in RT-PCR and Northern hybridization.
These kits provide a fast, reliable and convenient method to sequentially isolate and concentrate exosomal RNA as well as Free-Circulating RNA from different plasma/serum sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA as well as all sizes of the free-circulating protein-bound RNA, including microRNA. These kits provide a clear advantage over other available kits in that they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, these kits allow the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. Moreover, the free-circulating, protein-bound, RNA is free from any exosomal RNA. The purified RNA can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.
Plasma/Serum Exosome and Free-Circulating RNA Isolation Mini Kit
For sample volumes ranging from 50 µL to 1 mL.
Plasma/Serum Exosome and Free-Circulating RNA Isolation Midi Kit
For sample volumes ranging from 1 mL to 4 mL.
Plasma/Serum Exosome and Free-Circulating RNA Isolation Maxi Kit
For sample volumes ranging from 4 mL to 10 mL.
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| Kit Specifications | |
| Minimum Plasma Input | 1 mL |
| Maximum Plasma Input | 4 mL |
| Size of RNA Purified | All sizes, including miRNA and small RNA (< 200 nt) |
| Elution Volume | 50 – 100 µL |
| Time to Complete 10 Purifications | 40 – 45 minutes |
| Average Yields* | Variable depending on specimen |
*Please check page 5 of the product insert for the average yields and the common RNA quantification methods
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
Important Note
This kit is suitable for the purification of exosomes from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.
| Component | Cat. 59500 (50 preps) | Cat. 59600 (25 preps) | Cat. 59700 (15 preps) |
|---|---|---|---|
| Slurry E | 12.5 mL | 12.5 mL | 12.5 mL |
| ExoC Buffer | 8 mL | 8 mL | 2 x 8 mL |
| ExoR Buffer | 12 mL | 12 mL | 12 mL |
| Lysis Buffer A | 2 x 20 mL | 2 x 20 mL | 2 x 20 mL |
| Lysis Additive B | 2 mL | 2 mL | 2 mL |
| Wash Solution A | 2 x 18 mL | 18 mL | 18 mL |
| Elution Solution A | 2 x 6 mL | 6 mL | 6 mL |
| Mini Filter Spin Columns | 50 | 25 | 15 |
| Mini Spin Columns | 100 | 50 | 30 |
| Collection Tubes | 100 | 50 | 30 |
| Elution Tubes (1.7 mL) | 100 | 50 | 30 |
| Product Insert | 1 | 1 | 1 |